snoRNA Vectors for Sequence-specific Gene Knockdown in human cells

Genes carry the genetic instructions to allow cells to build proteins that control chemical reactions within cells and that regulate cell growth and division. Mutations can cause genes to make defective proteins that in turn cause many forms of human disease, including cancer, diabetes and inherited disorders. This proposal will develop a new technology that allows researchers to control which genes are active in human cells and also allows them to inactivate the defective copies of mutated genes and replace them with new genes that can now make the correct and active form of the same protein. This can be widely used to help test and develop new drugs and may also be developed as a direct therapy for use in patients.

This proposal aims to develop, optimize and evaluate a novel technology for the sequence-specific knock-down and replacement of one or multiple targeted genes in mammalian cells. This will provide a new approach for analysing gene function and for evaluating the effects of chemical inhibitors and drugs in human cells. The technology is based upon our recent discovery of a class of human small nucleolar RNAs (Box C/D snoRNAs) that can be targeted to inhibit the expression of cellular mRNAs and their protein products via the introduction of short regions of sequence complementarity within the snoRNA. This is the subject of a UK patent application entitled ?Targeted Modulation of Gene Expression?. The technology promises major advantages over existing methods, including the ability to knock-down two or more targeted genes simultaneously, thereby overcoming problems associated with analysing biological mechanisms involving genetic redundancy. It also provides a way to rapidly knock down one or more targeted genes and simultaneously replace them with a tagged and/or mutated version of the same gene, using a single vector.